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. 2017 May 3;7:1416. doi: 10.1038/s41598-017-01641-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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HAND2/TBX5 interaction drastically inhibited by Thalidomide. Protein-protein docking interface between TBX5 and HAND2 modeled DNA binding domain as retrieved from PDB. Only Residues of TBX5 within 5 angstroms of this interaction are shown (A). Those in red are associated with both TBX5-DNA and TBX5-HAND2 interaction from modeling simulation, which shows the position of thalidomide in the context of predicted TBX5-HAND2 interaction (B): TBX5 (cyan) and HAND2 (magenta). The physical interaction between both proteins was assessed by co-immunoprecipitation in the presence or absence (−) of either 4 µM thalidomide or DMSO (C). Ten times the quantity of proteins loaded for western blot was used for immunoprecipitation. Nuclear lysates of both HAND2 and TBX5 were immunoprecipitated with anti‐HA antibody and HAND2 proteins were visualized by western blot with and anti‐flag antibody. Membrane stripping and subsequent western blot analysis was performed with anti‐HA antibody in order to detect TBX5 proteins. Quantitation of the bands showed more than 80% decrease in the interaction when the extracts were incubated in the presence of Thalidomide. (Ctr: control, WB: immunoblot, IP: immnuoprecipitation).